Molecular characterization of irinotecan (SN-38) resistant human breast cancer cell lines

Haatisha Jandu; Kristina Aluzaite; Louise Fogh; Sebastian Wingaard Thrane; Julie B. Noer; Joanna Proszek; Khoa Nguyen Do; Stine Ninel Hansen; Britt Damsgaard; Signe Lykke Nielsen; Magnus Stougaard; Birgitta R. Knudsen; Jose Moreira; Petra Hamerlik; Madhavsai Gajjar; Marcel Smid; John Martens; John Foekens; Yves Pommier; Nils Brunner; Anne-Sofie Schrohl; Jan Stenvang

doi:10.1186/s12885-016-2071-1

Molecular characterization of irinotecan (SN-38) resistant human breast cancer cell lines

Haatisha Jandu, Kristina Aluzaite, Louise Fogh, Sebastian Wingaard Thrane, Julie B. Noer, Joanna Proszek, Khoa Nguyen Do, Stine Ninel Hansen, Britt Damsgaard, Signe Lykke Nielsen, Magnus Stougaard, Birgitta R. Knudsen, Jose Moreira, Petra Hamerlik, Madhavsai Gajjar, Marcel Smid, John Martens, John Foekens, Yves Pommier, Nils BrunnerAnne-Sofie Schrohl, Jan Stenvang

Research output: Contribution to journal › Journal article › Research › peer-review

744 Downloads (Pure)

Abstract

Background: Studies in taxane and/or anthracycline refractory metastatic breast cancer (mBC) patients have shown approximately 30 % response rates to irinotecan. Hence, a significant number of patients will experience irinotecan-induced side effects without obtaining any benefit. The aim of this study was to lay the groundwork for development of predictive biomarkers for irinotecan treatment in BC.Methods: We established BC cell lines with acquired or de novo resistance to SN-38, by exposing the human BC cell lines MCF 7 and MDA MB 231 to either stepwise increasing concentrations over 6 months or an initial high dose of SN-38 (the active metabolite of irinotecan), respectively. The resistant cell lines were analyzed for cross-resistance to other anti-cancer drugs, global gene expression, growth rates, TOP1 and TOP2A gene copy numbers and protein expression, and inhibition of the breast cancer resistance protein (ABCG2/BCRP) drug efflux pump.Results: We found that the resistant cell lines showed 7-100 fold increased resistance to SN-38 but remained sensitive to docetaxel and the non-camptothecin Top1 inhibitor LMP400. The resistant cell lines were characterized by Top1 down-regulation, changed isoelectric points of Top1 and reduced growth rates. The gene and protein expression of ABCG2/BCRP was up-regulated in the resistant sub-lines and functional assays revealed BCRP as a key mediator of SN-38 resistance.Conclusions: Based on our preclinical results, we suggest analyzing the predictive value of the BCRP in breast cancer patients scheduled for irinotecan treatment. Moreover, LMP400 should be tested in a clinical setting in breast cancer patients with resistance to irinotecan.

Original language	English
Article number	34
Journal	B M C Cancer
Volume	16
Issue number	1
Number of pages	13
ISSN	1471-2407
DOIs	https://doi.org/10.1186/s12885-016-2071-1
Publication status	Published - 2016

Keywords

ABCG2/BCRP
Breast cancer
Irinotecan
Resistance
SN-38
Topoisomerase I

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1186/s12885-016-2071-1

Art 3A10Final published version, 1.93 MB

OpenUrl availability

Full text

Cite this

Jandu, H., Aluzaite, K., Fogh, L., Thrane, S. W., Noer, J. B., Proszek, J., Nguyen Do, K., Hansen, S. N., Damsgaard, B., Nielsen, S. L., Stougaard, M., Knudsen, B. R., Moreira, J., Hamerlik, P., Gajjar, M., Smid, M., Martens, J., Foekens, J., Pommier, Y., ... Stenvang, J. (2016). Molecular characterization of irinotecan (SN-38) resistant human breast cancer cell lines. B M C Cancer, 16(1), Article 34. https://doi.org/10.1186/s12885-016-2071-1

@article{cba2ecdb23064426b06f2e29d4548fa0,

title = "Molecular characterization of irinotecan (SN-38) resistant human breast cancer cell lines",

abstract = "Background: Studies in taxane and/or anthracycline refractory metastatic breast cancer (mBC) patients have shown approximately 30 % response rates to irinotecan. Hence, a significant number of patients will experience irinotecan-induced side effects without obtaining any benefit. The aim of this study was to lay the groundwork for development of predictive biomarkers for irinotecan treatment in BC.Methods: We established BC cell lines with acquired or de novo resistance to SN-38, by exposing the human BC cell lines MCF 7 and MDA MB 231 to either stepwise increasing concentrations over 6 months or an initial high dose of SN-38 (the active metabolite of irinotecan), respectively. The resistant cell lines were analyzed for cross-resistance to other anti-cancer drugs, global gene expression, growth rates, TOP1 and TOP2A gene copy numbers and protein expression, and inhibition of the breast cancer resistance protein (ABCG2/BCRP) drug efflux pump.Results: We found that the resistant cell lines showed 7-100 fold increased resistance to SN-38 but remained sensitive to docetaxel and the non-camptothecin Top1 inhibitor LMP400. The resistant cell lines were characterized by Top1 down-regulation, changed isoelectric points of Top1 and reduced growth rates. The gene and protein expression of ABCG2/BCRP was up-regulated in the resistant sub-lines and functional assays revealed BCRP as a key mediator of SN-38 resistance.Conclusions: Based on our preclinical results, we suggest analyzing the predictive value of the BCRP in breast cancer patients scheduled for irinotecan treatment. Moreover, LMP400 should be tested in a clinical setting in breast cancer patients with resistance to irinotecan.",

keywords = "ABCG2/BCRP, Breast cancer, Irinotecan, Resistance, SN-38, Topoisomerase I",

author = "Haatisha Jandu and Kristina Aluzaite and Louise Fogh and Thrane, {Sebastian Wingaard} and Noer, {Julie B.} and Joanna Proszek and {Nguyen Do}, Khoa and Hansen, {Stine Ninel} and Britt Damsgaard and Nielsen, {Signe Lykke} and Magnus Stougaard and Knudsen, {Birgitta R.} and Jose Moreira and Petra Hamerlik and Madhavsai Gajjar and Marcel Smid and John Martens and John Foekens and Yves Pommier and Nils Brunner and Anne-Sofie Schrohl and Jan Stenvang",

year = "2016",

doi = "10.1186/s12885-016-2071-1",

language = "English",

volume = "16",

journal = "B M C Cancer",

issn = "1471-2407",

publisher = "BioMed Central Ltd.",

number = "1",

}

Jandu, H, Aluzaite, K, Fogh, L, Thrane, SW, Noer, JB, Proszek, J, Nguyen Do, K, Hansen, SN, Damsgaard, B, Nielsen, SL, Stougaard, M, Knudsen, BR, Moreira, J, Hamerlik, P, Gajjar, M, Smid, M, Martens, J, Foekens, J, Pommier, Y, Brunner, N, Schrohl, A-S & Stenvang, J 2016, 'Molecular characterization of irinotecan (SN-38) resistant human breast cancer cell lines', B M C Cancer, vol. 16, no. 1, 34. https://doi.org/10.1186/s12885-016-2071-1

TY - JOUR

T1 - Molecular characterization of irinotecan (SN-38) resistant human breast cancer cell lines

AU - Jandu, Haatisha

AU - Aluzaite, Kristina

AU - Fogh, Louise

AU - Thrane, Sebastian Wingaard

AU - Noer, Julie B.

AU - Proszek, Joanna

AU - Nguyen Do, Khoa

AU - Hansen, Stine Ninel

AU - Damsgaard, Britt

AU - Nielsen, Signe Lykke

AU - Stougaard, Magnus

AU - Knudsen, Birgitta R.

AU - Moreira, Jose

AU - Hamerlik, Petra

AU - Gajjar, Madhavsai

AU - Smid, Marcel

AU - Martens, John

AU - Foekens, John

AU - Pommier, Yves

AU - Brunner, Nils

AU - Schrohl, Anne-Sofie

AU - Stenvang, Jan

PY - 2016

Y1 - 2016

N2 - Background: Studies in taxane and/or anthracycline refractory metastatic breast cancer (mBC) patients have shown approximately 30 % response rates to irinotecan. Hence, a significant number of patients will experience irinotecan-induced side effects without obtaining any benefit. The aim of this study was to lay the groundwork for development of predictive biomarkers for irinotecan treatment in BC.Methods: We established BC cell lines with acquired or de novo resistance to SN-38, by exposing the human BC cell lines MCF 7 and MDA MB 231 to either stepwise increasing concentrations over 6 months or an initial high dose of SN-38 (the active metabolite of irinotecan), respectively. The resistant cell lines were analyzed for cross-resistance to other anti-cancer drugs, global gene expression, growth rates, TOP1 and TOP2A gene copy numbers and protein expression, and inhibition of the breast cancer resistance protein (ABCG2/BCRP) drug efflux pump.Results: We found that the resistant cell lines showed 7-100 fold increased resistance to SN-38 but remained sensitive to docetaxel and the non-camptothecin Top1 inhibitor LMP400. The resistant cell lines were characterized by Top1 down-regulation, changed isoelectric points of Top1 and reduced growth rates. The gene and protein expression of ABCG2/BCRP was up-regulated in the resistant sub-lines and functional assays revealed BCRP as a key mediator of SN-38 resistance.Conclusions: Based on our preclinical results, we suggest analyzing the predictive value of the BCRP in breast cancer patients scheduled for irinotecan treatment. Moreover, LMP400 should be tested in a clinical setting in breast cancer patients with resistance to irinotecan.

AB - Background: Studies in taxane and/or anthracycline refractory metastatic breast cancer (mBC) patients have shown approximately 30 % response rates to irinotecan. Hence, a significant number of patients will experience irinotecan-induced side effects without obtaining any benefit. The aim of this study was to lay the groundwork for development of predictive biomarkers for irinotecan treatment in BC.Methods: We established BC cell lines with acquired or de novo resistance to SN-38, by exposing the human BC cell lines MCF 7 and MDA MB 231 to either stepwise increasing concentrations over 6 months or an initial high dose of SN-38 (the active metabolite of irinotecan), respectively. The resistant cell lines were analyzed for cross-resistance to other anti-cancer drugs, global gene expression, growth rates, TOP1 and TOP2A gene copy numbers and protein expression, and inhibition of the breast cancer resistance protein (ABCG2/BCRP) drug efflux pump.Results: We found that the resistant cell lines showed 7-100 fold increased resistance to SN-38 but remained sensitive to docetaxel and the non-camptothecin Top1 inhibitor LMP400. The resistant cell lines were characterized by Top1 down-regulation, changed isoelectric points of Top1 and reduced growth rates. The gene and protein expression of ABCG2/BCRP was up-regulated in the resistant sub-lines and functional assays revealed BCRP as a key mediator of SN-38 resistance.Conclusions: Based on our preclinical results, we suggest analyzing the predictive value of the BCRP in breast cancer patients scheduled for irinotecan treatment. Moreover, LMP400 should be tested in a clinical setting in breast cancer patients with resistance to irinotecan.

KW - ABCG2/BCRP

KW - Breast cancer

KW - Irinotecan

KW - Resistance

KW - SN-38

KW - Topoisomerase I

U2 - 10.1186/s12885-016-2071-1

DO - 10.1186/s12885-016-2071-1

M3 - Journal article

C2 - 26801902

SN - 1471-2407

VL - 16

JO - B M C Cancer

JF - B M C Cancer

IS - 1

M1 - 34

ER -

Molecular characterization of irinotecan (SN-38) resistant human breast cancer cell lines

Abstract

Keywords

UN SDGs

Access to Document

OpenUrl availability

Fingerprint

Cite this